Rhodopin Incorporated into the Allochromatium vinosum LH2 Complex Is Able to Generate Singlet Oxygen under Illumination

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Abstract

DPA membranes from Allochromatium vinosum cells, in which carotenoid biosynthesis was inhibited using diphenylamine (DPA) were obtained, into which rhodopin was incorporated. The LH2 complex with rhodopin content of 85% was isolated. Using a test for the thermal stability of LH2 complexes (DPA and with incorporated rhodopin), it was established that carotenoids of the early stages of biosynthesis (≤1 molecules per complex) did not interfere with rhodopin incorporation. It was found that when the LH2 complex with incorporated rhodopin was irradiated with light at the wavelength of 502 nm, BChl850 was photobleached at a rate close to that in the control LH2 complex. This indicates that rhodopin, after being incorporated into the DPA LH2 complex, is capable of generating singlet oxygen under illumination. Previously obtained data on heterogeneity of the carotenoid composition in DPA LH2 complexes (variation in the number of individual carotenoids molecules per complex in the general population) and our earlier suggestion about the structural role of carotenoids, namely, their ability to stabilize the LH2 complexes, were confirmed. Based on analysis of our results, as well as of the literature data, the interaction of singlet oxygen and carotenoids is discussed.

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Z. K. Makhneva

Institute of Basic Biological Problems, Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Author for correspondence.
Email: andrey-moskalenko@rambler.ru
Russian Federation, Pushchino

M. A. Bolshakov

Institute of Basic Biological Problems, Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Email: andrey-moskalenko@rambler.ru
Russian Federation, Pushchino

A. A. Ashikhmin

Institute of Basic Biological Problems, Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Email: andrey-moskalenko@rambler.ru
Russian Federation, Pushchino

A. A. Moskalenko

Institute of Basic Biological Problems, Pushchino Scientific Center for Biological Research, Russian Academy of Sciences

Email: andrey-moskalenko@rambler.ru
Russian Federation, Pushchino

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2. Fig. 1. a - Absorption spectrum of rhodopin used for embedding (1) and LED emission spectrum; b - Absorption spectra of DFA membranes from cells of Alc. vinosum with inhibited Kar fusion before (1) and after addition of 3 portions of rhodopin to them and subsequent dialysis (2)

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3. Fig. 2. DFA absorption spectra of the LH2 complex (1), the LH2 complex with incorporated rhodopin (2), and the control LH2 complex from Alc. vinosum (3)

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4. Fig. 3. HPLC of pigments of the LH2 complex with integrated rhodopin (a) and DFA of the LH2 complex (b) from Alc. vinosum. Peak identification: 1-5 - BHL and its derivatives; 6 - didehydrorhodopin; 7 - rhodopin; 8 - spiriloxanthin; 9 - bacteriopheophytin; 10 - unidentified product of non-carotenoid nature; 11 - ζ-carotene; 12 - phytofluene; 13 - phytoin

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5. Fig. 4. Changes in the absorption spectrum of the LH2 complex with incorporated rhodopin (a) before (1) and after heating for 5 min at 80°C (2) and the DFA complex of LH2 (b) before (1) and after heating for 2 min (2) and 5 min (3) at 80°C. Spectra (c, d) were normalised: by maximum at 486 nm (c, 1) and at 590 nm (d, 1). Inset: PAGE of the DFA complex LH2 before (1) and after heating for 5 min (2) at 80°C (LH2 - LH2 complex; FP - free pigment zone)

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6. Fig. 5. Fading curves of BChl850 in DFA membranes (1), LH2 DFA complex (2), control LH2 complex (3) and LH2 complex with integrated rhodopin (4) under 502 nm LED illumination. Inset: difference spectra of ‘control - 30 min of illumination with 502 nm light’ of DFA-membranes (1) and DFA-complex LH2 (2). The spectra were normalised by the maximum of BChl850

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7. Fig. 6. Changes in the absorption spectrum of the LH2 complex with integrated rhodopin (a) and control LH2 complex (b) under illumination with an LED with an emission maximum of 502 nm: 1 - 0; 2 - 5; 3 - 10; 4 - 15; 5 - 20; 6 - 30 min

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8. Fig. 7. Effect of illumination with LED with emission maximum of 502 nm on LH2 complex with embedded rhodopin: a - increase of AtsCl absorption (1 - 5; 2 - 10; 3 - 15; 4 - 20; 5 - 30 min of illumination); b - absorption change curves of Qy BHL850 (1) and AtsCl (2)

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